Parental effects on offspring sex ratio in the Numbat (Myrmecobius fasciatus): does captivity influence paternal sex allocation?

Sex allocation theories predict that under different ecological conditions the production of sons and daughters will affect parental fitness differently. Skewed offspring sex ratios often occur under captive conditions where individuals are exposed to nutritional and social conditions that differ from nature. Here, we analyzed 29 years of offspring sex ratio data from a captive population of an endangered marsupial, the Numbat (Myrmecobius fasciatus). We partitioned variation in offspring sex ratio based on parental origin (captive- vs. wild-bred), parental weight, maternal age, and maternal reproductive history. Our analyses revealed no effect of parental weight or maternal origin on offspring sex ratio-however, there was a significant effect of paternal origin. Data visualization indicated that captive-bred males tended to produce male-biased litters. We discuss the result in relation to recent studies that have shown that male mammals have the capacity to be arbiters of sex allocation and highlight candidate mechanisms, but consider it with caution due to the small sample size from which the result was derived. We performed a population viability analysis (PVA) to explore the potential impact of a sex ratio skew on the sustainability of the captive Numbat population under hypothetical scenarios. Our PVA revealed that supplementation with wild individuals is critical to the persistence of the captive Numbat population and that a biased sex ratio will lead to extinction of the captive colony under certain conditions. Overall, our study demonstrates that covert sex ratio skews can persist undetected in captive populations, which have the potential to become impactful and compromise population sustainability under changed management processes.

Accelerometry reveals limits to use of an energy-saving anthropogenic food source by a threatened species: A case of Carnaby's cockatoos (Zanda latirostris) and canola.

The use of anthropogenic resources is becoming increasingly common as species adapt to human-induced environmental changes, but their use can expose species to new risks. Understanding how animals exploit these resources is important for guiding conservation management, particularly where species are threatened. The introduction of canola cropping to breeding areas of endangered Carnaby's cockatoo (Zanda latirostris) has been attributed to an increase in the birds' reproductive success; however, the seed may be protein-limiting for nestling growth and its use by cockatoos has been implicated in the emergence of a new disease. We used high-resolution accelerometer-capable GPS tags to track eight birds. Accelerometer data were used to calculate overall dynamic body acceleration (ODBA), a proxy for energy expenditure, and to identify and quantify canola and native vegetation foraging behaviours. We used linear mixed models to determine which factors affected patterns of resource use and to determine whether, and to what extent, canola use was associated with reduced energetic and movement costs. We then compared the energetic content of canola seed and native food sources to inform patterns of behaviour and habitat use revealed by our tracking data. Use of canola was associated with reduced movement costs and energy expenditure. However, there was an apparent reluctance to increase foraging on canola above a threshold of time, even when conditions reduced time available to utilise native food sources. While anthropogenic resources may appear to improve population trends in some cases, careful investigations of patterns of resource use are necessary to guide appropriate conservation management efforts. For Carnaby's cockatoos, conservation efforts should focus on retention, protection and expansion of native food sources.

Modeling genetic benefits and financial costs of integrating biobanking into the conservation breeding of managed marsupials.

Managed breeding programs are an important tool in marsupial conservation efforts but may be costly and have adverse genetic effects in unavoidably small captive colonies. Biobanking and assisted reproductive technologies (ARTs) could help overcome these challenges, but further demonstration of their potential is required to improve uptake. We used genetic and economic models to examine whether supplementing hypothetical captive populations of dibblers (Parantechinus apicalis) and numbats (Myrmecobius fasciatus) with biobanked founder sperm through ARTs could reduce inbreeding, lower required colony sizes, and reduce program costs. We also asked practitioners of the black-footed ferret (Mustela nigripes) captive recovery program to complete a questionnaire to examine the resources and model species research pathways required to develop an optimized biobanking protocol in the black-footed ferret. We used data from this questionnaire to devise similar costed research pathways for Australian marsupials. With biobanking and assisted reproduction, inbreeding was reduced on average by between 80% and 98%, colony sizes were on average 99% smaller, and program costs were 69- to 83-fold lower. Integrating biobanking made long-standing captive genetic retention targets possible in marsupials (90% source population heterozygosity for a minimum of 100 years) within realistic cost frameworks. Lessons from the use of biobanking technology that contributed to the recovery of the black-footed ferret include the importance of adequate research funding (US$4.2 million), extensive partnerships that provide access to facilities and equipment, colony animals, appropriate research model species, and professional and technical staff required to address knowledge gaps to deliver an optimized biobanking protocol. Applied research investment of A$133 million across marsupial research pathways could deliver biobanking protocols for 15 of Australia's most at-risk marsupial species and 7 model species. The technical expertise and ex situ facilities exist to emulate the success of the black-footed ferret recovery program in threatened marsupials using these research pathways. All that is needed now for significant and cost-effective conservation gains is greater investment by policy makers in marsupial ARTs.

Los programas de reproducción controlada son una herramienta importante para los esfuerzos de conservación de marsupiales, aunque pueden resultar costosos y tener efectos genéticos adversos en las colonias cautivas incapaces de aumentar en tamaño. Los biobancos y las tecnologías de reproducción asistida (TRA) podrían ayudar a superar estos problemas, pero es necesario seguir demostrando su potencial para mejorar su adopción. Utilizamos modelos genéticos y económicos para analizar si la introducción de esperma fundador proveniente de biobancos mediante tecnologías de reproducción asistida a poblaciones cautivas hipotéticas de los marsupiales Parantechinus apicalis y Myrmecobius fasciatus podría reducir la endogamia, disminuir el tamaño efectivo de las colonias y reducir el costo de los programas. También pedimos a los profesionales del programa de recuperación en cautiverio del hurón de patas negras (Mustella nigripes) que respondieran un cuestionario para analizar los recursos y los métodos de investigación de las especies modelo necesarias para desarrollar un protocolo de biobanco optimizado para el hurón de patas negras. Utilizamos los datos de este cuestionario para diseñar métodos de investigación con costos similares para los marsupiales australianos. Con el biobanco y la reproducción asistida, la endogamia se redujo en promedio entre un 80 y un 98%, el tamaño de las colonias fue en promedio un 99% más pequeño y los costos del programa entre 69 y 83 veces menores. La integración del biobanco posibilitó los objetivos de retención genética en cautiverio a largo plazo en marsupiales (90% de heterocigosidad de la población de origen durante un mínimo de 100 años) dentro de un marco realista de costos. Entre el aprendizaje extraído del uso de la tecnología de biobancos que contribuyó a la recuperación del hurón de patas negras figuran la importancia de una financiación adecuada de la investigación (4.2 millones de dólares), colaboraciones profundas que faciliten el acceso a instalaciones y equipos, colonias de animales, especies modelo adecuadas para la investigación y el personal profesional y técnico necesario para abordar las lagunas de conocimiento y ofrecer un protocolo optimizado para los biobancos. Una inversión en investigación aplicada de 133 millones de dólares australianos para la investigación de los marsupiales podría proporcionar protocolos de biobancos para 15 de las especies de marsupiales australianos en mayor riesgo y 7 especies modelo. Existen los conocimientos técnicos y las instalaciones ex situ para emular el éxito del programa de recuperación del hurón de patas negras en marsupiales amenazados utilizando estas vías de investigación. Ahora sólo se necesita una mayor inversión por parte de los responsables políticos de las TRA para marsupiales para obtener beneficios de conservación significativos y rentables.

An Efficient Method for the Euthanasia of Cane Toads (Rhinella marina) under Northern Australian Field Conditions.

The euthanasia of cane toads under field conditions presents a number of logistical and animal welfare challenges. One recommended method of control involves the use of carbon dioxide in plastic bags. This paper describes the minimum amount of time (4 h) required to efficiently euthanase toads with a carbon dioxide concentration of 4.96% under field conditions experienced in northern Australia. Discussion is also provided on the issues of safe disposal of biological and plastic waste associated with the application of this method.

Integrating biobanking could produce significant cost benefits and minimise inbreeding for Australian amphibian captive breeding programs.

Captive breeding is an important tool for amphibian conservation despite high economic costs and deleterious genetic effects of sustained captivity and unavoidably small colony sizes. Integration of biobanking and assisted reproductive technologies (ARTs) could provide solutions to these challenges, but is rarely used due to lack of recognition of the potential benefits and clear policy direction. Here we present compelling genetic and economic arguments to integrate biobanking and ARTs into captive breeding programs using modelled captive populations of two Australian threatened frogs, namely the orange-bellied frog Geocrinia vitellina and the white bellied frog Geocrinia alba . Back-crossing with frozen founder spermatozoa using ARTs every generation minimises rates of inbreeding and provides considerable reductions in colony size and program costs compared with conventional captive management. Biobanking could allow captive institutions to meet or exceed longstanding genetic retention targets (90% of source population heterozygosity over 100 years). We provide a broad policy direction that could make biobanking technology a practical reality across Australia's ex situ management of amphibians in current and future holdings. Incorporating biobanking technology widely across this network could deliver outcomes by maintaining high levels of source population genetic diversity and freeing economic resources to develop ex situ programs for a greater number of threatened amphibian species.

Measuring the Welfare Impact of Soft-Catch Leg-Hold Trapping for Feral Cats on Non-Target By-Catch.

To inform trapping protocols to reduce by-catch while trapping feral cats by-catch welfare costs should be quantified. During cat trapping programs at six Western Australian sites from 1997-2015, 431 non-target individuals, including 232 individuals from native species (132 mammals, 52 birds and 42 reptiles) were captured. Among the native fauna; birds were more likely to be severely injured (33%, compared to 12% in mammals and 21% in reptiles). Amongst other vertebrates, larger individuals were less likely to be injured. Olfactory lures used in these studies attracted reptiles, but repelled mammals. By-catch varied with climate and landscape. Trap injury to by-catch species poses ethical concerns, especially for threatened species that can least afford an additional threat. Future trapping should consider the timing of trapping, trap placement, trap settings (especially the treadle pressure needed to close the trap) and new innovations sending immediate capture alerts to minimise by-catch and potential injury associated with prolonged restraint. By-catch welfare data should be analysed to identify best practice and on-going improvement.

Finding the Balance: Fertility Control for the Management of Fragmented Populations of a Threatened Rock-Wallaby Species.

Populations of Australian marsupials can become overabundant, resulting in detrimental impacts on the environment. For example, the threatened black-flanked rock-wallaby ( Petrogale lateralis lateralis ) has previously been perceived as overabundant and thus 'unwanted' when they graze crops and cause habitat degradation. Hormonally-induced fertility control has been increasingly used to manage population size in other marsupials where alternative management options are not viable. We tested whether deslorelin, a superagonist of gonadotropin-releasing hormone (GnRH), would suppress reproduction in free-living adult female rock-wallabies without adversely impacting body condition. We trapped, synchronised reproduction and allocated female rock-wallabies to a placebo implant (control, n = 22), one (n = 22) or two (n = 20) subcutaneous implants of deslorelin. Females were then recaptured over the following 36 months to monitor reproduction, including Luteinising Hormone levels, and body condition. Following treatment, diapaused blastocysts reactivated in five females and the resulting young were carried through to weaning. No wallabies treated with deslorelin, conceivede a new young for at least 27 months. We did not observe adverse effects on body condition on treated females. We conclude that deslorelin implants are effective for the medium-term suppression of reproduction in female black-flanked rock-wallabies and for managing overabundant populations of some marsupials.

Reproductive implications of exposure to Toxoplasma gondii and Neospora caninum in western grey kangaroos (Macropus fuliginosus ocydromus).

Australian marsupials are thought to be particularly vulnerable to pathologic impacts of Toxoplasma gondii, and they may be similarly affected by Neospora caninum. Pathology due to either organism could be expressed as reduced female reproductive performance. We studied adult female western grey kangaroos (Macropus fuliginosus ocydromus) from suburban Perth, Western Australia, between May 2006 and October 2008. We used indirect fluorescent antibody tests to look for evidence of exposure to T. gondii and N. caninum in M. fuliginosus ocydromus and tested the association between their reproductive performance and a positive test result. Although 20% of plasma samples collected from 102 female kangaroos were positive for T. gondii and 18% were positive for N. caninum, we found no association between positive results and reproductive performance. Further study will be required to clarify if, and under what circumstances, T. gondii and N. caninum are pathogenic to macropod marsupials.

One fledgling or two in the endangered Carnaby's Cockatoo (Calyptorhynchus latirostris): a strategy for survival or legacy from a bygone era?

Of the five species of black cockatoo in the genus Calyptorhynchus, those species with red tail bands (Red-tailed Black Cockatoo and Glossy Black Cockatoo) lay clutches of only one egg and those with white or yellow tail bands (Carnaby's Cockatoo, Baudin's Cockatoo and Yellow-tailed Black Cockatoo) usually lay clutches of two. The breeding of the endangered Carnaby's Cockatoo has been studied from 1969 to 2012 at a number of localities throughout its range in south-western Australia within a region largely cleared for agriculture. When raising nestlings the species feeds on seeds of native vegetation, and there was a strong but not significant negative relationship between nesting success and percentage loss of native vegetation within 6 and 12 km of nest hollows. There was a significant negative relationship between the health of nestlings and percentage loss of native vegetation around nest hollows. While the usual clutch size is two, average clutch size tended to be lower in areas where much native vegetation has been cleared. While both eggs hatch in 77% of two-egg clutches, the species normally fledges only one young. However, the species is capable of fledging both nestlings from a breeding attempt. Sets of siblings are usually the product of older, more experienced females nesting in areas where more native vegetation has been retained. The conservation implications of these findings are discussed in the light of predicted changes to the climate of south-western Australia.

Application of STR markers in wildlife forensic casework involving Australian black-cockatoos (Calyptorhynchus spp.).

Parrots and cockatoos are highly prized aviary birds and the demands for such species has fuelled their illegal trade and harvest from the wild. Here we report on three forensic case studies involving black-cockatoos (Calyptorhynchus spp.) endemic to Australia. These cases involve suspected poaching and illegal killing of endangered red- and white-tailed black-cockatoos. Through the prior development of 20 polymorphic microsatellite loci and population databases for white- and red-tailed black-cockatoos, the tools are available to conduct high-resolution paternity and individual identity testing. In one case, we matched a red-tailed black-cockatoo nestling to a tree hollow from which it was poached through the use of DNA from eggshell recovered from the nest. For the second case, we utilized our provenance population database (nest sites), and identified the kinship and geographic origin of a white-tailed black-cockatoo, which was illegally harvested from the wild. The third case determined the number individual white-tailed black-cockatoos allegedly shot at a fruit grower's orchard from body part remains. These genetic investigations highlight the significance and statistical confidence of DNA profiling and associated databases for endangered taxa, such as exotic birds. Our cockatoo population databases are the first of their kind in Australia, and demonstrate the efficacy of such approaches to identify such illegal activity. With a robust set of genetic markers and methodologies in place, we aim to broaden our population databases to include other cockatoo species of conservation concern.

The evolutionary history of cockatoos (Aves: Psittaciformes: Cacatuidae).

Cockatoos are the distinctive family Cacatuidae, a major lineage of the order of parrots (Psittaciformes) and distributed throughout the Australasian region of the world. However, the evolutionary history of cockatoos is not well understood. We investigated the phylogeny of cockatoos based on three mitochondrial and three nuclear DNA genes obtained from 16 of 21 species of Cacatuidae. In addition, five novel mitochondrial genomes were used to estimate time of divergence and our estimates indicate Cacatuidae diverged from Psittacidae approximately 40.7 million years ago (95% CI 51.6-30.3 Ma) during the Eocene. Our data shows Cacatuidae began to diversify approximately 27.9 Ma (95% CI 38.1-18.3 Ma) during the Oligocene. The early to middle Miocene (20-10 Ma) was a significant period in the evolution of modern Australian environments and vegetation, in which a transformation from mainly mesic to xeric habitats (e.g., fire-adapted sclerophyll vegetation and grasslands) occurred. We hypothesize that this environmental transformation was a driving force behind the diversification of cockatoos. A detailed multi-locus molecular phylogeny enabled us to resolve the phylogenetic placements of the Palm Cockatoo (Probosciger aterrimus), Galah (Eolophus roseicapillus), Gang-gang Cockatoo (Callocephalon fimbriatum) and Cockatiel (Nymphicus hollandicus), which have historically been difficult to place within Cacatuidae. When the molecular evidence is analysed in concert with morphology, it is clear that many of the cockatoo species' diagnostic phenotypic traits such as plumage colour, body size, wing shape and bill morphology have evolved in parallel or convergently across lineages.